Latent TGF-β2 Structure and Activation

潜在 TGF-β2 结构和激活

基本信息

批准号：
10446300
负责人：
TIMOTHY A SPRINGER
金额：
$ 70.65万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10446300
关键词：
2019-nCoV Actins Affinity Antibodies Aortic Aneurysm Binding Biochemical Biological Biological Process Blood Vessels Cardiovascular Physiology Cardiovascular system Cell Line Cell surface Cells Chimeric Proteins Complex Cryoelectron Microscopy Crystallization Crystallography Cytoplasmic Tail Cytoskeleton Defect Detection Development Disease Extracellular Matrix Family Follow-Up Studies Foundations Grant Growth Factor Heart Human Integral Membrane Protein Integrin Binding Integrins Knowledge LRRC32 gene Length Loeys-Dietz Syndrome Mediating Membrane Modeling Molecular Chaperones Molecular Conformation Mucocutaneous Lymph Node Syndrome Mus Mutagenesis Mutate Mutation N-terminal Patients Peptide Hydrolases Peptides Physiological Pilot Projects Preparation Process Proteins RGD (sequence)Rana Reagent Research Role Scleroderma Serine Protease Site Structure Systemic Scleroderma Testing Therapeutic Time Tissues Transforming Growth Factor Beta 2 Transforming Growth Factor beta Transforming Growth Factor beta Receptors Vertebrates Virus X-Ray Crystallography cardiac repair cofactor coronary fibrosis dimer disulfide bond extracellular latent TGF-beta binding protein member monomer nanobodies novel novel therapeutics prevent receptor repaired therapeutic target

项目摘要

Abstract. Transforming growth factor β2 (TGF-β2) is critically important for heart and vascular development and repair. TGF-β2 dysregulation is seen in patient TGF-β2 mutations, systemic sclerosis, and Kawasaki disease, which have cardiovascular sequelae such as aortic aneurysms and cardiac fibrosis. TGF-β1, 2 and 3 are synthesized as proproteins that dimerize and associate with milieu molecules that regulate TGF-β tissue localization, such as the transmembrane protein glycoprotein A repetitions predominant (GARP) and latent TGF-β binding proteins (LTBPs) in the extracellular matrix (ECM). Proconvertases cleave between the prodomain and growth factor (GF) domain; however, the prodomain dimer remains non-covalently associated with the GF in a proTGF-β–milieu molecule complex after secretion. ProTGF-β–milieu molecule complexes are inactive because the prodomains encircle the GF and prevent binding to TGF-β receptors. ProTGF-β1 and 3 activation is mediated by binding of integrins αVβ6 and αVβ8 to an RGD-motif in the prodomain and requires proTGF-β association with a milieu molecule. How proTGF-β2, which lacks an RGD-motif, is activated remains a mystery. Aim 1 will define the structure of proTGF-β2 to understand its mechanism of latency. Aim 2 will determine proTGF-β2/milieu molecule complex structures by X-ray crystallography and cryo-EM to define how milieu molecules bind and alter TGF-β2 latency. We will generate antibodies to use as crystallization chaperones in addition to using already developed nanobodies to proTGF-β2. Complementary unfolding studies will test the hypothesis that milieu molecule binding stabilizes proTGF-β2. Aim 3 characterizes TGF-β2 activation. Follow-up studies will identify cell-lines that natively activate TGF-β2 and characterize the physiologically relevant process. The results of this grant will enhance our understanding of TGF-β2 latency and activation in extracellular milieus and lay the foundation for developing therapeutics that target proTGF-β2 and its physiologically relevant complexes with milieu molecules.

抽象的。转化生长因子β2（TGF-β2）对于心脏和血管发育至关重要和维修。 TGF-β2失调在患者TGF-β2突变，全身性硬化和川崎中可见疾病，患有心血管后遗症，例如主动脉瘤和心脏纤维化。 TGF-β1、2和3 合成为原则，与调节TGF-β组织的环境分子二聚并缔合定位，例如跨膜蛋白糖蛋白A重复（GARP）和潜在细胞外基质（ECM）中的TGF-β结合蛋白（LTBP）。 Proconvertases在 Prodomain和生长因子（GF）域；但是，prodomain二聚体仍然无共价关联分泌后的ProtgF-β-Milieu分子复合物中的GF。 Protgf-β-Milieu分子络合物是无活性是因为该蛋白元环绕GF并防止与TGF-β受体结合。 Protgf-β1和3 通过整联蛋白αVβ6和αVβ8与Prodomain中的RGD-MOTIF结合，激活是介导的，需要 Protgf-β与环境分子的关联。缺少RGD-MOTIF的ProtgF-β2如何被激活一个神秘的。 AIM 1将定义ProtgF-β2的结构，以了解其潜伏期的机理。 AIM 2意志通过X射线晶体学和冷冻EM确定ProtgF-β2/Milieu分子复合物结构，以定义如何定义环境分子结合并改变TGF-β2潜伏期。我们将生成用于结晶的抗体除了使用已经开发的ProtgF-β2的纳米词外，伴侣。互补的发展研究将检验以下假设：环境分子结合稳定ProtgF-β2。目标3个字符TGF-β2 激活。后续研究将鉴定出本质地激活TGF-β2并表征的细胞耳。生理上相关的过程。该赠款的结果将增强我们对TGF-β2潜伏期的理解和细胞外环境中的激活，并为靶向Protgf-β2的治疗奠定基础及其与环境分子的物理相关复合物。